Human Isogenic Organoid Models of Genetic Forms of Autism to Identify Convergent and Divergent Pathomechanisms in Autism

自闭症遗传形式的人类同基因类器官模型，用于识别自闭症趋同和发散的病理机制

• 批准号: 10736309
• 负责人: Stefan Aigner
• 金额: $ 79万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-07 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10736309
• 关键词: 3-Dimensional; Acceleration; Address; Affect; Architecture; Autopsy; Awareness; Biological Assay; Brain; Calcium; Categories; Cell Line; Cell model; Cells; Child; Chromium; Chromosome Mapping; Clinical; Complex; Copy Number Polymorphism; Corpus striatum structure; Data; Data Set; Defect; Development; Diagnostic; Disease model; Enzymes; Etiology; FRAP1 gene; Frequencies; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Models; Genetic Transcription; Genome; Genome engineering; Human; Image; Individual; Interneurons; Investigation; Measures; Messenger RNA; Methodology; Methods; Modeling; Molecular; Nature; Neurodevelopmental Disorder; Neurons; Noise; Operative Surgical Procedures; Organoids; Pathologic; Pathology; Pathway interactions; Patients; Phenotype; Population; Prosencephalon; Protein Biosynthesis; Proteins; RNA Editing; Recurrence; Regulation; Reproducibility; Research Personnel; Resolution; Resources; Ribosomal Interaction; Ribosomal Proteins; Ribosomes; Role; Series; Signal Transduction; Sirolimus; Site; Synapses; Testing; Tissues; Translations; Variant; apoB mRNA editing catalytic subunit; autism spectrum disorder; biomarker discovery; biomarker identification; brain tissue; candidate validation; cell type; diagnostic biomarker; disorder risk; drug testing; functional outcomes; induced pluripotent stem cell; insight; microdeletion; migration; multi-electrode arrays; neuronal circuitry; pharmacologic; predictive signature; rabies viral tracing; risk variant; single-cell RNA sequencing; stable cell line; stem cell model; stem cells; targeted treatment; therapeutic development; tool; transcriptome; transcriptomics; transgene expression; translatome

PROJECT SUMMARY Autism spectrum disorder (ASD) is a clinically complex, heterogeneous condition affecting 1 in 44 children in the U.S. The identification of common etiologies across multiple forms of genetic and idiopathic forms of ASD will critically advance diagnostic biomarker discovery and therapeutic development. Dysregulation of cellular translation has emerged as a pathophysiological mechanism common to at least a subset of ASD forms. However, systematic investigation of the cellular mechanisms that converge onto the ASD phenotype has been hampered by a paucity of robust and reproducible human ASD cellular models and scalable experimental tools for cell-type resolved characterization at the level of translation. To address these bottlenecks and to directly address the role of translational dysregulation as a common feature in ASD, we have (1) used advanced genome engineering tools to generate an extensively validated, isogenic series of induced pluripotent stem cell (iPSC) lines modeling 15 syndromic forms of ASD caused by highly penetrant gene and genome variants, representing ~10% of the total ASD population (the largest such panel created to date, to our knowledge), (2) established a robust human iPSCs-derived cortical organoid model of brain development, and (3) developed ribo-STAMP, a method for translational profiling of individual cells in heterogeneous cell populations, which is the first and only method enabling translation to be measured at single- cell resolution. In this project, we identify common and divergent pathological mechanisms in genome- engineered isogenic stem cell based organoid models of ASD, using single-cell transcriptomic and translatomic approaches. We validate our findings using cellular and functional phenotypic assays and in patient-derived iPSC models. If successful, our study will identify common and unique translation-aware single-cell resolved gene expression signatures that predict cellular and functional outcomes. We anticipate that our datasets and insights into cell-type specific deficits in gene expression of genetic forms of autism will critically accelerate the development of a unified framework that enables molecular categorization of both genetic and idiopathic cases, facilitating the identification of biomarkers and the development of targeted therapies.

项目摘要 自闭症谱系障碍（ASD）是一种临床上复杂的异质疾病，影响44名儿童中有1个 美国识别跨多种遗传和特发性ASD形式的常见病因将 严重提高诊断生物标志物发现和治疗性开发。细胞失调 翻译已成为至少一部分ASD形式共有的病理生理机制。 然而，对收敛到ASD表型的细胞机制的系统研究已经 由于鲁棒和可重复可重复的人ASD细胞模型和可扩展的实验工具的阻碍 用于细胞类型在翻译水平上的分解表征。解决这些瓶颈并直接 解决转化失调作为ASD中常见特征的作用，我们有（1）使用高级基因组 工程工具生成了经过广泛验证的，等生的诱导多能干细胞（IPSC） 模拟高度渗透基因和基因组变体引起的15种综合征形式的ASD综合征形式的线，代表 约有10％的ASD总人口（据我们所知，迄今为止创建的最大的此类小组），（2）建立了一个 强大的人IPSC衍生的皮质器官大脑发育模型，（3）开发了Ribo-Stamp，A 在异质细胞种群中单个细胞翻译分析的方法，这是第一个也是唯一的 可以在单细胞分辨率下测量翻译的方法。在这个项目中，我们确定了常见和 基因组工程性等源性干细胞基于ASD的基因组基因器官模型的不同病理机制， 使用单细胞转录组和翻译方法。我们使用细胞和 功能表型测定和患者衍生的IPSC模型。如果成功，我们的研究将确定常见 以及独特的翻译感知单细胞分辨基因表达特征，这些基因表达信号预测细胞和功能 结果。我们预计我们的数据集和对细胞类型的基因表达中特定缺陷的见解 自闭症的遗传形式将批判性地加速统一框架的发展 遗传和特发性病例的分类，促进生物标志物和 靶向疗法的开发。